Aralkyl-N-hydroxyureas as inhibitors of 5-lipoxygenase and oxidation of low density lipoprotein

ABSTRACT

This invention relates to compounds compounds useful in treating diseases mediated by one or more leukotrienes or oxidative modification of low density lipoprotein such as inflammation, bronchoconstriction or atherosclerosis. The compounds of this invention have the formula: ##STR1## wherein R 1  and R 3  are independently hydrogen, fluorine, chlorine, bromine, iodine, C 1  -C 6  alkyl, trifluoromethyl, C 1  -C 6  alkoxy, or C 1  -C 6  trifluoroalkoxy; R 2  and R 4  are hydrogen or methyl independently; 
     R 5  is hydrogen, methyl or hydroxy; 
     X and Z are independently oxygen or sulfur; 
     and 
     Y is --CH 2  --, --CH(CH 3 )--, or --CH═CHCH(CH 3 )--.

This invention relates to aralkyl-N-hydroxyureas which have5-lipoxygenase inhibition properties, a method for inhibiting thebiosynthesis of leukotrienes and pharmaceutical compositions forinhibiting leukotriene synthesis and are thus useful as antiinflammatoryand anti-asthma agents. Compounds of this invention also have theability to scavenge peroxyl radicals implicated in the oxidation of lowdensity lipoprotein and thus are useful in the treatment of conditionsfor which the inhibition of oxidative modification of lipids isindicated such as atherosclerosis.

BACKGROUND OF THE INVENTION

Leukotrienes participate in inflammatory reactions, exhibit chemotacticactivities, stimulate lysosomal enzyme release and act as importantfactors in the intermediate hypersensitivity reaction. Leukotrienes aremetabolic products of metabolism of arachidonic acid (AA) bylipoxygenase enzymes with the most significant leukotrienes being LTB₄,LTC₄, LTD₄ and LTE₄. The latter three leukotrienes are incorporated inthe substance known as SRS or SRS-A, the slow-reacting substance ofanaphylaxis [J. Immun. 215, 115-118 (1980), Biochem. Biophys. Res. Comm.93, 1121-1126(1980)]. By another metabolic pathway, arachidonic acid ismetabolized by cyclooxygenase enzymes to prostaglandins andthromboxanes.

Leukotrienes LTC₄ and LTD₄ are potent bronchoconstrictors of the humanbronchi [Dahlen et al., Nature 288,484-486 (1980) and Piper, Int. Arch.Appl, Immunology 76, Suppl. 1, 43 (1985)] which stimulate the release ofmucus from airways in vitro [Macom et al., Am. Rev. Resp. Dis. 126, 449(1982)], are potent vasodilators in skin [Bisgaard et al.,Prostaglandins 23, 797 (1982)], and produce a wheal and flare response[Camp et al., Brit. J. Pharmacol. 80 497 (1983)]. The nonpeptideleukotriene LTB₄ is a powerful chemotactic factor for leukocytes [A. S.Ford - Hutchinson, J. Royal Soc. Med. 74, 831-883 (1981)], whichstimulates cell accumulation and affects vascular smooth muscle [Bray,Brit. Med. Bull. 39, 249 (1983)]. The activity of leukotrienes asmediators of inflammation and hypersensitivity is extensively reviewed[Bailey and Casey, Ann. Reports Med. Chem. 17, 203-217 (1982) and Bray,Agents and Actions 19, 87 (1986) and Masamune and Melvin, Ann. ReportsMed. Chem. 24, 71 (1989)].

There is also evidence that products of the cyclooxygenase/lipoxygenasepathways play key roles in both the pathogenesis of gastric mucosaldamage due to extracellular (gastric and intestinal contents,microorganisms, and the like) or intracellular (ischemia, viruses, etc.)agents, as well as in cytoprotection against such damage. Thus, on theone hand prostaglandins exert a cytoprotective effect on the gastricmucosa [see Robert, Gastroenterology, 77, 761-767 (1979)] and thisaction of the prostaglandins, especially of the E series, is consideredto be of importance in the treatment of gastrointestinal ulceration [seeIsselbacher, Drugs, 33 (suppl.), 38-46 (1987)]. On the other hand, exvivo experiments have shown that gastric mucosal tissue fromethanol-pretreated rats is capable of LTC₄ generation and that this LTC₄production is quantitatively related to the severity of the ethanoldamage [see Lange et al., Naunyn-Schmiedeberg's Arch, Pharmacol. Suppl.,330, R27, (1985)]. It has also been demonstrated that LTC₄ can inducevasoconstriction in both venous and arteriolar vessels in the ratsubmucosa [see Whittle, IUPHAR Ninth Int. Cong. of Pharmac., S30-2,London, England (1984)]. This is significant since ethanol-inducedlesion formation in gastric mucosa may be multifactorial with, forexample, stasis of gastric blood flow contributing significantly to thedevelopment of the hemorrhagic necrotic aspects of the tissue injury[see Guth et al., Gastroenterology, 87, 1083-90 (1984)]. Moreover, inthe anesthetized cat, exogenous LTD₄ evokes both increased pepsinsecretion and decreased transgastric potential [Pendleton et al., Eur.J. Pharmacol., 125, 297-99 (1986)]. A particularly significant recentfinding in this regard is that 5-lipoxygenase inhibitors and someleukotriene antagonists protect the gastric mucosa against lesionsinduced by the oral or parenteral administration of most nonsteroidalantiinflammatory drugs [see Rainsford, Agents and Actions, 21, 316-19(1987)]. Accordingly, a significant body of evidence implicates theinvolvement of lipoxygenase products in the development of pathologicalfeatures associated with gastric mucosal lesions, such as for examplethose induced by ethanol exposure and administration of non-steroidalanti-inflammatory drugs. Thus, compounds which inhibit the biologicaleffects of leukotrienes and/or which control the biosynthesis of thesesubstances, as by inhibiting 5-lipoxygenase, are considered to be ofvalue as cytoprotective agents.

Accordingly, the biological activity of the leukotrienes and SRS's, andof lipoxygenase as the enzyme leading to the metabolism of AA toleukotrienes, indicates that a rational approach to drug therapy toprevent, remove or ameliorate the symptoms of allergies, anaphylaxis,asthma and inflammation and for gastric cytoprotection must focus oneither blocking the release of mediators of these conditions orantagonizing their effects. Thus compounds, which inhibit the biologicaleffects of the leukotrienes and SRS's and/or which control thebiosynthesis of these substances, as by inhibiting lipoxygenase, areconsidered to be of value in treating such conditions as allergicbronchial asthma, allergic rhinitis, as well as in other immediatehypersensitivity reactions and in providing gastric cytoprotection.

Compounds of this invention inhibit lipoxygenase and antagonize productsof the lipoxygenase pathway and thus are useful as antiinflammatory andanti-allergic agents. Compounds of this invention are expected to havegastric cytoprotective activity.

Atherosclerosis, the underlying disease implicated in myocardialinfarction and strokes, is a complex pathologic process involving theintimal layer of the arteries. The earliest lesion of atherosclerosis isdevelopment of the fatty streak lesions which contain lipid-ladenmacrophages and lipid-laden smooth muscle cells. Macrophages do not takeup native low density lipoprotein (LDL) but do take up modified, i.e.,acetylated LDL or oxidized LDL via acetyl-LDL or "scavenger" receptorsto form the foam cells of atherosclerotic plaque. Free radial oxidation,i.e., lipid peroxidation, has been shown to be involved in thealteration of LDL by endothelial cells. Arterial smooth muscle cellsgenerate superoxide and oxidize LDL in the presence of micromolarconcentrations of Cu⁺² or Fe⁺². The way LDL can be modified byendothelial cells can be mimicked in vitro by incubation of thelipoprotein in the presence of CuCl₂. Probucol, an antihyperlipidemicagent, also inhibits both cell mediated and Cu⁺² mediated oxidativemodification of LDL, and was shown to inhibit the formation ofatherosclerotic lesions in WHHL rabbits [Reaven et al., Arteriosclerosisand Thrombosis 12(3), 318-324 (1992), Steinberg, The Amer. J. ofCardiology 57, 16H-21H (1986), Carew. Schwenke and Steinberg, Proc.Natl. Acad. Sci. 84, 7725-7729 (1987) and Nagano et al.,Arteriosclerosis 9 (4), 453-461 (1989)]. Thus in vitro inhibition ofCu⁺² catalyzed oxidation of LDL is indicative of antiatheroscleroticutility.

Compounds of this invention inhibit in vitro the copper-inducedperoxidation of LDL and thus are useful in the treatment or preventionof arteriosclerosis.

Lipoxygenase inhibiting compounds of the formula: ##STR2## wherein R¹ isamino or methyl, R² is C₁ -C₂ alkyl, R³ is selected from hydrogen,halogen, and trihalomethyl; R⁴ is hydrogen, halogen, trihalomethyl, C₁-C₄ alkoxy or C₁ -C₄ alkyl; and M is hydrogen, a pharmaceuticallyacceptable cation, aroyl or C₁ -C₆ alkoyl are disclosed in EP 279281 A2.EP 0384594 A1 discloses antiinflammatory 5-lipoxygenase inhibitors ofthe formula: ##STR3## wherein R¹ and R² are selected from H and C₁ -C₄alkyl independently and the 4-halophenoxy moiety can be attached to thephenyl ring at either the 3 or 4 position.

The PCT patent WO 90/12008 (equivalent to U.S. Pat. No. 5,185,363)discloses lipoxygenase inhibiting compounds of the formula: ##STR4##wherein M is, among other choices, hydrogen or a pharmaceuticallyacceptable cation; Z is O or S; X is a straight or branched C₁ -C₆alkylene or C₂ -C₆ alkenylene group optionally substituted by hydroxy,halogen, cyano, alkoxy, aminocarbonyl, carboxy and alkoxycarbonyl; R¹and R² are independently hydrogen, hydroxy, or C₁ -C₆ alkyl optionallysubstituted by hydroxy, halogen, cyano, alkoxy, etc., with a provisothat both R¹ and R₂ cannot hydroxy; and R₃ can be phenyl, naphthyl orthienyl optionally substituted by a variety of substituents includingcarbocyclic or heterocyclic arylalkoxy groups optionally substituted byhalogen, nitro, cyano, alkyl, alkoxy or halosubstituted alkyl whereinthe heterocyclic aryl moiety is defined as a 5 to 6 membered ringcontaining one N, S, or O atom or a N and O or a N and S or three Natoms and further stipulates that the 5 to 6 membered heterocyclicarylmoiety may be fused with a phenyl ring to form a benzo-fusedheterocycle. The PCT application WO 92/03425 (equivalent to U.S. Pat.No. 5,334,604) discloses compounds that are intermediates toantidiabetic compounds having the formula: ##STR5## wherein n is 0 or 1,R is hydrogen or C₁ -C₃ alkyl, R³ is one of C₁ -C₉ alkyl, C₃ -C₇cycloalkyl, phenyl, naphthyl, furyl, benzofuryl or thienyl optionallysubstituted with one or two groups selected from C₁ -C₃ alkyl, C₁ -C₃alkoxy, C_(1-C) ₃ alkoxycarbonyl, trifluoromethyl, fluoro or chloro; R⁶is hydrogen, C₁ -C₉ alkyl, C₃ -C₇ cycloalkyl, phenyl, naphthyl, furyl,benzofuryl or thienyl and R⁷ is hydrogen or a conventional protectinggroup. N-Aryl-N-hydroxy ureas, formamides and alkylamides havingpreviously been disclosed to have lipoxygenase inhibiting activity, aredisclosed in WO 92/03130 as having anti-atherosclerotic activity andhave the formula:

    Ar--Y--Q

wherein Ar is heteroaromatic, naphthyl, tetrahydronaphthyl, phenyl orphenyl substituted by phenyl, naphthyl or a heteroaromatic group; Y isC₁ -C₁₀ alkylene or C₂ -C₁₀ alkenelene and Q is ##STR6## where R¹ is H,C₁ -C₄ alkyl or an Ar group and R² is H, C₁ -C₄ alkyl, amino, mono ordialkylamino, cycloalkylamino, cycloalkylalkylamino, anilino,N-alkylanilino or an Ar group.

SUMMARY OF THE INVENTION

The compounds useful in the methods and pharmaceutical compositions ofthis invention are represented by Formula I: ##STR7## wherein: R¹ and R³are independently hydrogen, fluorine, chlorine, bromine, iodine, C₁ -C₆alkyl, trifluoromethyl, C₁ -C₆ alkoxy, or C₁ -C₆ trifluoroalkoxy;

R² and R⁴ are independently hydrogen or methyl;

R⁵ is hydrogen, methyl, or hydroxy;

X and Z are oxygen or sulfur independently;

and

Y is --CH₂ --, --CH(CH₃)--, or --CH═CHCH(CH₃)--.

By way of further definition the term C₁ -C₆ trifluoroalkoxy denotes aC₁ -C₆ alkoxy group in which the hydrogens of the terminal methyl groupare replaced by fluorines, and is preferably a trifluoromethoxy ortrifluoroethoxy group.

It is an object of this invention to provide novel compounds of FormulaI. Another object of this invention is to provide methods of treatingdiseases of inflammation, asthma and atherosclerosis in which synthesesof leukotrienes by lipoxygenase and oxidation of lipoprotein, especiallylow density lipoprotein (LDL) are inhibited. Still another object is toprovide pharmaceutical compositions for the treatment of inflammation,asthma or atherosclerosis.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of the invention can be prepared according to thefollowing representative synthetic schemes. The variables R¹ -R⁵, X andZ are as defined above. Other variables are as defined hereinbelow.

a. When Y is --CHR⁶ -- where R⁶ is H or methyl; the invention compoundsare prepared as indicated in Scheme I. ##STR8##

b. When Y is --CH═CH--CH(CH₃)--, the invention compounds can be preparedaccording to the synthetic reaction sequence shown in Scheme II.##STR9##

c. When R⁴ or R⁵ independently hydrogen and hydroxy or methyl andhydroxy, the invention compounds can be prepared according to syntheticreaction schemes III and IV. ##STR10##

The intermediate 2-phenyl-4-chloromethyl-5-(H or methyl) oxazoles andthiazoles can be prepared by known methods conventional in the art(Heterocyclic Compounds 34, 1979 and Heterocyclic Compounds 45, 1986)The 2-phenyl-4-chloromethyl-5-methyloxazoles can be prepared accordingto the reaction sequence shown in Scheme V. ##STR11##

The intermediate 4-chloromethyl-2-phenyloxazoles or thiazoles can beprepared according to the reaction shown in Scheme VI. ##STR12## X=O orS

The 1,3-bis(tetrahydropyran-2-yloxy)urea used in Schemes III and IV canbe prepared by reaction of the known O-tetrahydropyran-2-ylhydroxylamine (Warrener and Cain, Angew, Chem, Int. Ed. 5, 511 (1955))with 1,1'-carbonyldiimidazole.

The following specific examples are included for illustrative purposesonly and are not to be construed as limiting to the scope of theinvention. Other synthetic procedures may be apparent to those skilledin the art. In the following examples, reagents and intermediates areeither commercially available or can be prepared according to standardliterature procedures by those skilled in the art.

EXAMPLE 11-Hydroxy-1-[3-(5-methyl-2-phenyl-oxazol-4-ylmethoxy)-benzyl]-urea

Step a) 3-(5-Methyl-2-phenyl-oxazol-4-ylmethoxy)-benzaldehyde

A mixture of 4-chloromethyl-5-methyl-2-phenyl-oxazole (8.25 g, 39.7mmol), 3-hydroxybenzaldehyde (4.85 g, 39.7 mmol), potassium carbonate(5.49 g, 39.7 mmol) and dimethylformamide (100 mL) was stirred at 80° C.for 8 hours. The mixture was then poured into H₂ O and extracted withEtOAc. The organic extracts were dried over MgSO₄. Evaporation andpurification by flash chromatography on silica gel (eluting solventEtOAc/hexane 4/1), gave a yellow solid (10.5 g, 90% yield, m.p.104°-105° C.).

Analysis for: C₃₈ H₃₆ N₂ O₉ Calc'd: C, 73.71; H, 5.15; N, 4.78 Found: C,73.88; H, 5.10; N, 4.66

Step b) 3-(5-Methyl-2-phenyl-oxazol-4-ylmethoxy)-benzaldehyde oxime

In to a solution of3-(5-methyl-2-phenyl-oxazol-4-ylmethoxy)-benzaldehyde (10.0 g, 34.1mmol), in ethanol (400 mL) were added hydroxylamine hydrochloride (7.11g, 102.39 mmol) and a solution of sodium acetate (11.19 g, 136.52 mmol)in H₂ O (40 mL). The mixture was stirred at room temperature for 10hours, then poured into H₂ O, acidified with 2N HCl and extracted withEtOAc. The organic extracts were dried over MgSO₄. Evaporation andcrystallization from acetone/ether/hexane, gave a white solid (8.95 g,85% yield, m.p. 163°-164° C.)

Analysis for: C₁₈ H₁₆ N₂ O₃ Calc'd: C, 70.12; H, 5.23; N, 9.09 Found: C,70.01; H, 5.23; N, 8.90

Step c)N-[3-(5-methyl-2-phenyl-oxazol-4-ylmethoxy)-benzyl]-hydroxylamine

In to a solution of3-(5-methyl-2-phenyl-oxazol-4-ylmethoxy)-benzaldehyde oxime (8.0 g, 25.9mmol) in MeOH (400 mL) and THF (80 mL) were added sodiumcyanoborohydride (8.06.9 g, 129.5 mmol) and methyl orange (indicator, 20mg). A solution of 4N HCl in dioxane was then added dropwise in order tomaintain a pH range 3 to 4. When a persistent red color was observed thereaction mixture was poured into H₂ O, basified with 2N NaOH to pH about8 to 9 and extracted with EtOAc. The organic extracts were dried overMgSO₄. Evaporation and purification by flash chromatography, on silicagel (eluting solvent EtOAc/MeOH 10/1), gave a white solid (6.5 g, 81%yield, m.p. 123°-124° C.).

Analysis for: C₁₈ H₁₈ N₂ O₃ Calc'd: C, 69.66; H, 5.85; N, 9.03 Found: C,69.34; H, 5.88; N, 8.75

Step d)1-Hydroxy-1-[3-(5-methyl-2-phenyl-oxazol-4-ylmethoxy)benzyl]-urea

In to a solution ofN-[3-(5-methyl-2-phenyl-oxazol-4-ylmethoxy-benzyl]-hydroxylamine (1.25g, 4.03 mmol) in dioxane (20 mL) was added trimethylsilylisocyanate(0.85 mL, 605 mmol). The mixture was stirred at room temperature for 2hour, poured into H₂ O, acidified with 2N HCl and extracted with EtOAc.The organic extracts were dried over MgSO₄. Evaporation andcrystallization from acetone/ether, gave a white solid (0.98 g, 69%yield, m.p. 151°-152° C.).

Analysis for: C₁₉ H₁₉ N₃ O₄ Calc'd: C, 64.58; H, 5.42; N, 11.89 Found:C, 64.83; H, 5.54; N, 11.84

EXAMPLE 21-Hydroxy-1-[3-[5-methyl-2-(4-trifluoromethyl-phenyl)-oxazol-4-ylmethoxy]-benzyl]-urea

Step a)3-[5-Methyl-2-(4-trifluoromethyl-phenyl)-oxazol-4-ylmethoxy]-benzaldehyde

The title compound was prepared similar to the method described inexample 1, step a), and was obtained as a yellow solid in 65% yield,m.p. 104°-105° C.

Analysis for: C₁₉ H₁₄ NO₃ F₃ Calc'd: C, 63.16; H, 3.91; N, 3.88 Found:C, 62.84; H, 3.97; N, 3.87

Step b)3l-[5-Methyl-2-(4-trifluoromethyl-phenyl)-oxazol-4-ylmethoxy]-benzaldehydeoxime

The title compound was prepared similar to the method described inexample 1, step b), and was obtained as a yellowish solid in 62% yield,m.p. 125°-126° C.

Analysis for: C₁₉ H₁₅ F₃ N₂ O₃ Calc'd: C, 60.64; H, 4.02; N, 7.44 Found:C, 60.82; H, 4.04; N, 7.38

Step c)N-[3-[5-methyl-2-(4-trifluoromethyl-phenyl)-oxazol-4-ylmethoxy]-benzyl]-hydroxylamine

The title compound was prepared similar to the method described inexample 1, step c), and was obtained as an off-white solid in 76% yield.The product was carried to the next step without purification.

Step d)1-Hydroxy-1-[3-[5-methyl-2-4-(trifluoromethyl-phenyl)-oxazol-4-ylmethoxy]-benzyl)-urea

The title compound was prepared similar to the method described inexample 1, step d), and was obtained as a white solid, in 56% yield,m.p. 142°-143° C.

Analysis for: C₂₀ H₁₈ F₃ N₃ O₃ Calc'd: C, 57.01; H, 4.31; N, 9.97 Found:C, 57.10; H, 4.33; N, 9.60

EXAMPLE 31-[3-[2-(4-fluoro-phenyl)-oxazol-4-ylmethoxy]-benzyl]-1-hydroxy-urea

Step a) 3-[2-(4-fluoro-phenyl)-oxazol-4-ylmethoxy]benzaldehyde

The title compound was prepared similar to the method described inexample 1, step a), and was obtained as a solid in 80.0% yield, m.p.89°-91° C.

Analysis for: C₁₇ H₁₂ FNO₃ Calc'd: C, 68.68; H, 4.07; N, 4.71 Found: C,68.39; H, 3.89; N, 4.99

b) 3-[2-(4-fluoro-phenyl)-oxazol-4-ylmethoxy]benzaldehyde oxime

The title compound was prepared similar to the method descirbe inexample 1, step b) and was obtained as a white solid 92.0 % yield, m.p.144°-154° C. (decomposed).

Analysis for: C₁₇ H₁₃ FN₂ O₃ Calc'd: C, 65.38; H, 4.20; N, 8.97 Found:C, 65.14; H, 3.89; N, 4.99

step c)N-hydroxy-N-[3-[2-(4-fluoro-phenyl)-oxazol-4-ylmethoxy]-phenyl-1-ylmethyl]-amine

The title compound was prepared similar to the method described inexample 1, step c), and was obtained as a solid in 88.0% yield, m.p.115°-120° C. (decomposed), MS (EI, m/e): 314 (M)⁺.

step d)1-Hydroxy-1-[3-[2-(4-fluoro-phenyl)-oxazol-4-ylmethoxy]-benzyl]-urea

The title compound was prepared similar to the method described inexample 1, step d), and was obtained as a white solid in 71.0 % yield,m.p. 176°-177° C. (decomposed).

Analysis for: C₁₈ H₁₆ FN₃ O₄ Calc'd: C, 60.50; H, 4.51; N, 11.76 Found:C, 60.32; H, 4.43; N, 11.73

EXAMPLE 41-Hydroxy-1-[3-[2-(4-fluoro-phenyl)-oxazol-4-ylmethoxy]-benzyl]-thiourea

The title compound was prepared similar to the method described inexample 1, step d), except that trimethylsilylthioisocyanate is used inplace of trimethylsilylisocyanate. The product is obtained as a whitesolid in 61.0% yield, m.p. 188°-189° C. (decomposed).

Analysis for: C₁₈ H₁₆ FN₃ O₃ S Calc'd: C, 57.89; H, 4.32; N, 11.25Found: C, 58.05; H, 4.24; N, 11.45

EXAMPLE 5 1-Hydroxy-1-[3-(2-phenyl-thiazol-4-ylmethoxy)-benzyl]-urea

Step a) 3-(2-phenyl-thiazol-4-ylmethoxy)benzaldehyde

The title compound was prepared similar to the method described inexample 1, step a), was obtained as a solid in 91.0% yield, m.p. 68°-71° C.

Analysis for: C₁₇ H₁₃ NO₂ S Calc'd: C, 69.13; H, 4.44; N, 4.74 Found: C,68.99; H, 4.44; N, 4.76

Step b) 3-(2-phenyl-thiazol-4-ylmethoxy)benzaldehyde oxime

The title compound was prepared similar to the method described inexample 1, step b), was obtained as a solid in 86.0% yield, m.p.140°-144° C.

Analysis for: C₁₇ H₁₄ N₂ O₂ S Calc'd: C, 65.79; H, 4.55; N, 9.03 Found:C, 65.85; H, 4.52; N, 9.23

Step c)N-hydroxy-N-[3-(2-phenyl-thiazol-4-ylmethoxy)-phenyl-1-ylmethyl]-amine

The title compound was prepared similar to the method described inexample 1, step c) and was obtained as a solid in 98.0% yield, m.p.120°-128° C. (decomposed), MS (CI, m/e): 312 (M+H)⁺.

Step d) 1-Hydroxy-1-[3-(2-phenyl-thizaol-4-ylmethoxy)-benzyl]-urea

The title compound was prepared similar to the method described inexample 1, step d) and was obtained as a solid in 62.5% yield, m.p.158°-163° C.

Analysis for: C₁₈ H₁₇ N₃ O₃ S Calc'd: C, 60.83; H, 4.82; N, 11.82 Found:C, 60.87; H, 4.73; N, 11.83

EXAMPLE 61-Hydroxy-1-[3-[2-(4-chloro-phenyl)-thiazol-4-ylmethoxy]-benzyl]-urea

Step a) 3-[2-(4-chloro-phenyl)-thiazol-4-ylmethoxy]benzaldehyde

The title compound was prepared similar to the method described inexample 1, step a) and was obtained as a solid in 68% yield, m.p.119°-123° C.

Analysis for: C₁₇ H₁₂ ClNO₂ S Calc'd: C, 61.91; H, 3.67; N, 4.25 Found:C, 62.11; H, 3.53; N, 4.27

Step b) 3-[2-(4-chloro-phenyl)-thiazol-4-ylmethoxy]benzaldehyde oxime

The title compound was prepared similar to the method described inexample 1, step b) and was obtained as a white solid in 96.5% yield,m.p. 162°-171° C.

Analysis for: C₁₇ H₁₂ ClN₂ O₂ S Calc'd: C, 59.22; H, 3.80; N, 8.12Found: C, 59.57; H, 3.63; N, 8.11

Step c)N-hydroxy-N-[3[2-(4-chloro-phenyl)-thiazol-4-ylmethoxy]-phenyl-1-ylmethyl]-amine

The title compound was prepared similar to the method described inexample 1, step c), and was obtained as a solid in 95.1% yield, m.p.132°-133° C.

Analysis for: C₁₇ H₁₅ ClN₂ O₂ S Calc'd: C, 58.87; H, 4.36; N, 8.08Found: C, 58.43; H, 4.26; N, 7.91

Step d)1-Hydroxy-1-[3-[2-(4-chloro-phenyl)-thiazol-4-ylmethoxy]-benzyl]-urea

The title compound was prepared similar to the method described inexample 1, step d), and was obtained as a white solid in 68.3% yield,m.p. 164°-167° C. (decomposed).

Analysis for: C₁₈ H₁₆ ClN₃ O₃ S Calc'd: C, 55.45; H, 4.14; N, 10.78Found: C, 55.52; H, 4.08; N, 10.70

EXAMPLE 71-Hydroxy-1-[3-[2-(4-methoxy-phenyl)-thiazol-4-ylmethoxy]-benzyl]-urea

Step a) 3-[2-(4-methoxyphenyl)thiazol-4-ylmethoxy]benzaldehyde

The title compound was prepared similar to the method described inexample 1, step a) and was obtained as a solid in 91.0% yield, m.p.119°-121° C.

Analysis for: C₁₈ H₁₅ NO₃ S Calc'd: C, 66.44; H, 4.65; N, 4.30 Found: C,66.36; H, 4.60; N, 4.40

Step b) 3-[2-(4-methoxy-phenyl)-thiazol-4-ylmethoxy]benzaldehyde oxime

The title compound was prepared similar to the method described inexample 1, step b), and was obtained as a white solid in 99.0% yield,m.p. 144°-153° C. (decomposed).

Analysis for: C₁₈ H₁₆ ClN₂ O₂ S Calc'd: C, 63.51; H, 4.74; N, 8.23Found: C, 63.90; H, 4.65; N, 8.28

Step c)N-hydroxy-N-[3-[2-(4-methoxy-phenyl)-thiazol-4-ylmethoxy]-phenyl-1-ylmethyl]-amine

The title compound was prepared similar to the method described inexample 1, step c), and was obtained as a solid in 88.7% yield, m.p.132°-133° C. (decomposed), MS (El, m/e): 342 (M)⁺.

Step d)1-Hydroxy-1-[3-]2-(4-methoxy-phenyl)-thiazol-4-ylmethoxy]-benzyl]-urea

The title compound was prepared similar to the method described inexample 1, step d), and was obtained as a solid in 66.7% yield, m.p.179°-184° C. (decomposed), MS (EI, m/e): 342 (M)⁺.

Analysis for: C₁₉ H₁₉ N₃ O₄ S Calc'd: C, 59.21; H, 4.97; N, 10.90 Found:C, 59.38; H, 4.94; N, 10.80

EXAMPLE 81-[3-[2-(4-Chloro-phenyl)-5-methyl-oxazol-4-ylmethoxy]-benzyl]-1-hydroxy-urea

Step a) 3-[5-Methyl-2-(4-chlorophenyl)-oxazol-4-ylmethoxy]-benzaldehyde

The title compound was prepared similar to the method described inexample 1, step a) and was obtained as a white solid in 85% yield, m.p.115°-117° C.

Analysis for: C₁₈ H₁₄ ClNO₃ Calc'd: C, 65.96; H, 4.31; N, 4.24 Found: C,65.96; H, 4.20; N, 4.10

Step b) 3-[5-Methyl-2-(4-chlorophenyl)-oxazol-4-ylmethoxy]-benzaldehydeoxime

The title compound was prepared similar to the method described inexample 1, step b), and was obtained as a white solid in 96% yield, m.p.140°-142° C.

Analysis for: C₁₈ H₁₅ ClN₂ O₃ Calc'd: C, 63.07; H, 4.41; N, 8.17 Found:C, 62.96; H, 4.35; N, 8.05

Step c)N-[3-[5-methyl-2-(4-chlorophenyl)-oxazol-4-ylmethoxy]-benzyl]-hydroxylamine

The title compound was prepared similar to the method described inexample 1, step c), and was obtained as a white solid in 90% yield, m.p.125°-127° C.

Analysis for: C₁₈ H₇ ClN₂ O₃ Calc'd: C, 62.08; H, 4.97; N, 8.12 Found:C, 61.98; H, 4.85; N, 7.98

Step d)1-Hydroxy-1-[3-[2-(4-chloro-phenyl)-5-methyl-oxazol-4-ylmethoxy]benzyl]-urea

The title compound was prepared similar to the method described inexample 1, step d), and was obtained as a white solid in 69% yield, m.p.166°-167° C.

Analysis for: C₁₉ H₁₈ ClN₃ O₄ Calc'd: C, 58.84; H, 4.68; N, 10.84 Found:C, 58.64; H, 4.61; N, 10.70

EXAMPLE 91-[3-[2-(4-fluorophenyl)-5-methyl-oxazol-4-ylmethoxy]-benzyl]-1-hydroxy-urea

The title compound was prepared in substantially the same manner asdescribed in example 1, steps a-d, and was obtained as a white solid,m.p. 156°-157° C.

Analysis for: C₁₉ H₁₈ FN₃ O₄ Calc'd: C, 61.45; H, 4.89; N, 11.32 Found:C, 61.65; H, 4.88; N, 11.28

EXAMPLE 101-Hydroxy-1-[3-[2-(3-trifluoromethyl-phenyl)-5-methyl-oxazol-4-ylmethoxy]-benzyl]-urea

The title compound was prepared in substantially the same manner asdescribed in example 1, steps a-d, and was obtained as a white solid,m.p. 160°-161 ° C.

Analysis for: C₂₀ H₁₈ F₃ N₃ O₄ Calc'd: C, 57.01; H, 4.31; N, 9.97 Found:C, 57.07; H, 4.20; N, 9.64

EXAMPLE 111-[3-[2-(3,5-Bis-trifluoromethyl-phenyl)-5-methyl-oxazol-4-ylmethoxy]-benzyl]-1-hydroxy-urea

The title compound was prepared in substantially the same manner asdescribed in example 1, steps a-d, and was obtained as a white solid,m.p. 150°-151° C.

Analysis for: C₂₁ H₁₇ F₆ N₃ O₄ Calc'd: C, 51.53; H, 3.47; N, 8.59 Found:C, 51.56; H, 3.47; N, 8.41

EXAMPLE 121-[3-Fluoro-5-[5-methyl-2-(4-trifluoromethyl-phenyl)-oxazol-4-ylmethoxy]-benzyl]-1-hydroxy-urea

The title compound was prepared in substantially the same manner asdescribed in example 1, steps a-d, and was obtained as a white solid,m.p. 156°-157° C.

Analysis for: C₂₀ H₁₇ F₄ N₃ O₄ Calc'd: C, 50.25; H, 2.81; N, 3.26 Found:C, 50.11; H, 2.69; N, 3.16

EXAMPLE 131-[3-Fluoro-5-(5-methyl-2-phenyl-oxazol-4-ylmethoxy)-benzyl]-1-hydroxy-urea

The title compound was prepared in substantially the same manner asdescribed in example 1, steps a-d, and was obtained as a white solid,m.p. 157°-158° C.

Analysis for: C₁₉ H₁₈ FN₃ O₄ Calc'd: C, 61.45; H, 4.88; N, 11.31 Found:C, 61.11; H, 5.10; N, 11.13

EXAMPLE 141-Hydroxy-1-[1-[3-(5-methyl-2-phenyl-oxazol-4-ylmethoxy)-phenyl]-ethyl]-urea

The title compound was prepared in substantially the same manner asdescribed in example 1, steps a-d. 3'-Hydroxyacetophenone was used inplace of 3-hydroxybenzaldehyde. The title compound was obtained as awhite solid, m.p. 172°-173° C.

Analysis for: C₂₀ H₂₁ N₃ O₄ Calc'd: C, 65.38; H, 5.76; N, 11.44 Found:C, 65.11; H, 5.72; N, 11.19

EXAMPLE 15(E)-1-Hydroxy-1-[1-methyl-3-[3-(2-phenyl-thiazol-4-ylmethoxy)-phenyl]-allyl-urea

Step a ) 3-(2-phenyl-thiazol-4-ylmethoxy)-benzaldehyde

The title compound was prepared in substantially the same manner asdescribed in example 1, step a), and was obtained as a white solid in93% yield, m.p. 68°-70° C.

Analysis for: C₁₇ H₁₃ NO₂ S₂ Calc'd: C, 69.13; H, 4.43; N, 4.74 Found:C, 68.73; H, 4.25; N, 4.62

Step b) (E)-4-[3-(2-phenyl-thiazol-4-ylmethoxy)-phenyl]-but-3,en-2-one

In to a mixture of 3-(2-phenyl-thiazol-4-ylmethoxy)benzladehyde (10.0 g,33.89 mmol) and acetone (100 mL) was added dropwise a solution of NaOH(2.3 g, 57.6 mmol) in H₂ O (20 mL). After stirring for 1 hour the excessacetone was removed in vacuo and the residue was acidified with 1N HCl.After 30 minutes the mixture was extracted with EtOAc and the organicextracts were dried over MgSO₄. The product was carried to the next stepwithout further purification, MS (m/e): 335 (M⁺).

Step c) (E)-4-[3-(2-phenyl-thiazol-4-ylmethoxy)-phenyl]-but-3-en-2-ol

In to a cold (-20° C.) solution of(E)-4-[3-(2-phenyl-thiazol-4-ylmethoxy)-phenyl]-but-3-en-2-one (11.35 g,33.89 mmol) in THF (30 mL) and MeOH (20 mL) were added CeCl₃.7H₂ O (12.6g, 33.89 mmol), followed by portionwise addition of sodium borohydride(1.29 g, 33.89 mmol). After stirring for 30 minutes, the mixture waspoured into H₂ O, acidified with 2N HCl and extracted with EtOAc. Theorganic extracts were dried over MgSO₄. Evaporation and purification byflash chromatography on silica gel, (eluting solvent, hexane/EtOAc,3/1), gave a yellow solid (10.5 g, 92% yield, m.p. 75°-77° C.).

Analysis for: C₂₀ H₁₉ NO₂ S Calc'd: C, 71.19; H, 5.68; N, 4.15 Found: C,71.33; H, 5.64; N, 4.10

Step d)(E)-N-[3-[3-(2-phenyl-thiazol-4-ylmethoxy)-phenyl]-1-methyl-allyl]-hydroxylamine

In to a cold (-20° C.) mixture of(E)-4-[3-(2-phenyl-thiazol-4-ylmethoxy)-phenyl]-but-3-en-2-ol (3.7 g,11.0 mmol) tert-butyl-N-(tert-butoxy-carbonyloxy)carbamate (2.82 g,12.10 mmol, triphenylphosphine (3.17 g, 12.10 mmol) and THF (60 mL), wasadded dropwise a solution of diethylazodicarboxylate (1.9 mL, 12.10mmol) in THF (20 mL). The reaction mixture was allowed to come to 0° C.,stirred for 30 minutes, poured into H₂ O and extracted with EtOAc. Theorganic extracts were dried over MgSO₄. Evaporation gave a yellowviscous oil, which was taken up in methylene chloride (50 mL) andtreated with trifluoroacetic acid (20 mL). After stirring for 3 hoursthe volatiles were removed in vacuo and the residue was dissolved inEtOAc. The organic phase was washed with H₂ O, 1N NaOH and brine.Evaporation and purification by flash chromatogrtaphy on silica gel(eluting solvent EtOAc/MeOH, 10/1), gave a white solid (2.1 g, 54%yield, m.p. 78°-80° C.).

Analysis for: C₂₀ H₂₀ N₂ O₂ S Calc'd: C, 68.15; H, 5.72; N, 7.95 Found:C, 67.75; H, 5.74; N, 7.63

Step e)(E)-1-Hydroxy-1-[1-methyl-3-[3-(2-phenyl-thiazol-4-ylmethoxy)-phenyl]-allyl]-urea

The title compound was prepared similar to the method described inexample 1, step d), and was obtained as a white solid in 68% yield, m.p.152°-153° C.

Analysis for: C₂₁ H₂₁ N₃ O₃ S Calc'd: C, 63.78; H, 5.35; N, 10.63 Found:C, 63.94; H, 5.48; N, 10.41

EXAMPLE 16(E)-1-[3-[3-[2-(4-chlorophenyl)-5-methyl-oxazol-4-ylmethoxy]-phenyl]-1-methyl-allyl]-1-hydroxyl-urea

The title compound was prepared by substantially the same manner asdescribed in example 15, steps a-e), and was obtained as a white solid,m.p. 98°-99° C.

Analysis for: C₂₂ H₂₂ ClN₃ O Calc'd: C, 61.75; H, 5.18; N, 9.80 Found:C, 61.60; H, 5.17; N, 9.40

EXAMPLE 171,3-Dihydroxy-1-[4-(5-methyl-2-phenyl-oxazol-4-ylmethoxy)-benzyl]-urea

Step a) 3-(5-methyl-2-phenyl-oxazol-4-ylmethoxy)-phenyl-methanol

In to a solution of3-(5-methyl-2-phenyl-oxazol-4-ylmethoxy)-benzaldehyde (6.6 g, 22.52mmol) in MeOH (150 mL) was added portionwise sodium borohydride (845 mg,22.52 mmol). After stirring for two hours, the mixture was poured intoH₂ O, and extracted with EtOAc. The organic extracts were dried overMgSO₄. Evaporation gave a clear oil (6.2 g, 94% yield)

Analysis for: C₁₈ H₁₇ NO₃ Calc'd: C, 73.20; H, 5.80; N, 4.24 Found: C,72.73; H, 5.84; N, 4.59

Step b) 4-[3-(2-chloro-methyl)-phenoxymethyl]-5-methyl-2-phenyl-oxazole

In to a cold (0° C.) mixture of phosphorus pentachloride (4.23 g, 20.34mmol), CaCO₃ (2.03 g, 20.34 mmol) and ethyl ether (50 mL) was added asolution of 3-(5-methyl-2-phenyl-oxazol-4-ylmethoxy)phenyl-methanol (6.0g, 20.34 mmol) in THF (20 mL). After stirring for 30 minutes the mixturewas poured into H₂ O and extracted with ether. The organic extracts weredried over MgSO₄. Evaporation and purification by flash chromatographyon silica gel (eluting solvent EtOAc/hexane, 1/8) gave a yellowish oil(4.6 g, 72% yield). MS(m/e): 314 (M+H⁺).

Step c) 1,3-bis-(tetrahydro-pyran-2-yloxy)-urea

A mixture of tetrahydro-pyran-2-yl-hydroxylamine (5.0 g, 42.73 mmol),1,1'-carbonyldiimidazole (3.46 g, 21.37 mmol) and THF (20 mL) wasrefluxed for 10 hours. The mixture was poured into H₂ O and extractedwith EtOAc. The organic extracts were dried over MgSO₄. Evaporation andpurification by flash chromatography on silica gel (eluting solventhexane/EtOAc 1/1), gave a white solid (3.6 g, 32% yield, m.p. 98°-99°C.).

Analysis for: C₁₁ H₂₀ N₂ O₅ Calc'd: C, 50.76; H, 7.75; N, 10.76 Found:C, 50.91; H, 7.86; N, 10.78

Step d)1,3-dihydroxy-1-[4-(5-methyl-2-phenyl-oxazol-4-ylmethoxy)-benzyl]-urea

In to a mixture of 1,3-bis-(tetrahydro-pyran-2-yloxy)-urea (2.9 g, 11.16mmol) and DMF (15 mL) was added portionwise sodium hydride (80% inmineral oil, 335 mg, 11.16 mmol). After stirring for 1 hour,4-[3-(2-chloro-methyl)-phenoxymethyl]-5-methyl-2-phenyl-oxazole (3.5 g,11.16 mmol) was added and the mixture was stirred for 5 hours at roomtemperature. The mixture was then poured into H₂ O, acidified with 1NHCl, and extracted with EtOAc. The organic extracts were dried overMgSO₄. Evaporation gave a yellowish oil, which was taken up in MeOH (20mL) and treated with 4N HCl in dioxane (5 mL). After stirring at roomtemperature for 10 hours the mixture was poured into H₂ O and extractedwith EtOAc. The organic extracts were dried over MgSO₄. Evaporation andpurification by flash chromatography, on silica gel (eluting solventhexane/EtOAc 1/2), gave a white solid (1.21 g, 71% yield, m.p. 194°-195°C.).

Analysis for: C₁₉ H₁₉ N₃ O₅ Calc'd: C, 61.78; H, 5.18; N, 11.38 Found:C, 61.97; H, 5.33; N, 11.21

EXAMPLE 181,3-Dihydroxy-3-methyl-1-[3-(5-methyl-2-phenyl-oxazol-4-ylmethoxy)benzyl]-urea

Step a) 4-[3-(2-bromo-methyl)-phenoxymethyl]-5-methyl-2-phenyl-oxazole

A mixture of 2-[3-(5-methyl-2-phenyl-oxazol-4-ylmethoxy-phenyl]-methanol(6.0 g, 20.34 mmol), dioxane (20 mL) and HBr (48%, 50 mL) was stirred atroom temperature for 24 hours. The mixture was then poured into H₂ O andextracted with EtOAc. The organic extracts were dried over MgSO₄.Evaporation and purification by flash chromatography, on silica gel(eluting solvent hexane/EtOAc 5/1), gave a yellowish oil (5.8 g, 80%yield). MS (m/e): 357 (M⁺)

Step b) 1,3-bis-(tetrahydro-pyran-2-yloxy)-1-methyl-urea

In to a cold (0° C.) solution of 1,3-bis-(tetrahydro-pyran-yloxy)-urea(5.8 g, 22.32 mmol) in THF (50 mL) was added lithiumbis(trimethylsilyl)amide solution (22.32 mL, 22.32 mmol). After stirringfor 2 hours, methyl iodide (2.78 mL, 44.64 mmol) was added and themixture was stirred at room temperature for 10 hours. Then, the mixturewas poured into H₂ O, acidified with 1N HCl and extracted with EtOAc.The organic extracts were dried over MgSO₄. Evaporation and purificationby flash chromatography on silica gel (eluting solvent hexane/EtOAc 1/2)gave a white solid (4.6 g, 75% yield, m.p. 102°-104° C.).

Analysis for: C₁₂ H₂₂ N₂ O₅ Calc'd: C, 52.54; H, 8.08; N, 10.21 Found:C, 52.92; H, 8.12; N, 9.93

Step c)1,3-dihydroxy-3-methyl-1-[3-(5-methyl)-2-phenyl-oxazol-4-ylmethoxy)-benzyl]urea

In to a cold (0° C.) solution of1,3-bis-(tetrahydro-pyran-2-yloxy)-l-methyl-urea (2.52 g, 9.22 mmol) inTHF (20 mL) was added lithium bis(trimethylsilyl)amide solution (9.92ml, 9.92 mmol). The mixture was stirred for 1 hour, and4-[3-(2-bromo-methyl)-phenoxymethyl]-6-methyl-2-phenyl-oxazole (3.3 g,9.22 mmol) in THF (5 mL) was added dropwise. After stirring at roomtemperature for 5 hours, the mixture was poured into H₂ O, acidifiedwith 1N HCl and extracted with EtOAc. The organic extracts were driedover MgSO₄. Evaporation gave a yellowish oil, which was taken in MeOH(20 mL) and treated with 4N HCl in dioxane (5 mL). After stirring for 10hours, the mixture was poured into H₂ O and extracted with EtOAc. Theorganic extracts were dried over MgSO₄. Evaporation and purification byflash chromatography on silica gel (eluting solvent hexane/EtOAc 1/2)gave a white solid (1.26 g, 36 % yield, m.p. 62°-64° C).

Analysis for: C₂₀ H₂₁ N₃ O₅ Calc'd: C, 62.65; H, 5.52; N, 10.96 Found:C, 63.00; H, 5.69; N, 10.56

PHARMACOLOGY

Lipoxygenase inhibiting activity and inhibition of Cu⁺² mediatedperoxidation of LDL by the Formula I compounds is demonstrated inseveral standard pharmacological assays.

1. Inhibition of 5-Lipoxygenase in Human Whole Blood

Blood is obtained in 50-100 ml quantities from male donors. White bloodcell counts and differentials are made. Two ml of blood are placed in a15 ml polypropylene test tube. Test compounds are solubilized indimethylsulfoxide and diluted in 1:10 in 10% bovine serum albumin inphosphate buffered saline, pH 7.4 resulting in a final dimethylsulfoxideconcentration of 0.1% in the blood. Then, solutions of test compoundsare added to the blood in a shaking water bath at 37° C. for 10 minutesprior to the addition of 30 μM calcium ionophore (A23 187, Sigma). Afterionophore administration, whole blood samples are mixed and incubatedfor 20 minutes at 37° C. in a shaking water bath. Incubation isterminated by placing samples in an ice bath and immediately addingethylene glycol-bis-(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid (10mM). Samples are mixed and centrifuged at 1200 x g for 15 minutes at 4°C. Preparation of samples for evaluation by RIA or ELISA is carried outby the following protocol. Plasma is removed from sample tubes, placedin 15 ml polypropylene test tubes containing 8 ml methanol, and thenvortexed to precipitate protein. Samples are stored at -70° C.overnight. The next day, samples are centrifuged at 200 x g for 15minutes at 4° C. to pellet the precipitate. Samples are dried in aSavant SpeedVac Concentrator, Model SVC 200H, reconstituted to originalvolume with ice cold RIA or ELISA buffer, and stored at -70° C. untilassayed. The assay for eicosanoids (LTB₄, TxB₂, and PGE₂) is performedas described by the manufacturer of the [³ H]-RIA kit or ELISA kit (LTB₄-Amersham, TxB₂ and PGE₂ -Caymen Chemical).

The total eicosanoid level in 2 ml of blood is calculated and reportedas ng/10⁶ neutrophils. Significance is determined by a one-way analysisof variance with least significant difference (LSD) comparisons tocontrol (p≦0.05) and IC₅₀ 's (μM) are determined by regression analysis(Finney, 1978). Drug effects are expressed as percent change fromcontrol values. The results for compounds of this invention tested inthis assay are presented in Table I.

                  TABLE I                                                         ______________________________________                                        Compound of   Dose                                                            Example No.   (μM) % Inhibition of LTB.sub.4                               ______________________________________                                        1             1       52                                                      2             1       48                                                      3             1       74                                                      4             1       61                                                      5             1       66                                                      6             1       68                                                      7             1       50                                                      8             1       77                                                      9             1       74                                                      10            1       51                                                      11            10      57                                                      12            10      85                                                      13            1       70                                                      14            10      80                                                      15            10      47                                                      16            10      57                                                      17            10      83                                                      18            10      69                                                      ______________________________________                                    

2. Ex-Vivo Measurement of Lipoxygenase Inhibition in Orally Dosed Rats

Male Sprague Dawley rats (Charles River) weighing between 180 g and 200g were dosed orally with an invention compound (1-25 mg/kg, po)suspended in 0.5% Tween 80®. After an interval of 3 or 6 hours, the ratswere anesthetized with metofane (n=4/group) and blood collected inheparinized tubes. White blood cell counts and differentials were made.One ml of blood from each animal was placed in a 5 ml plastic tube in ashaker bath at 37° C. A23187 at a final concentration of 10 μM was addedto the blood and the blood vortexed and incubated for 15 min at 370° C.with gentle shaking. Incubation was terminated by vortexing sample andcentrifuging immediately at 1200 xG for 15 min at 4° C. The plasma wastransferred to 15 ml plastic tubes each containing 8 ml of methanol toprecipitate protein followed by vortexing. The samples were stored at-70 ° C. overnight and the next day the samples were centrifuged at 800xG for 15 min to pellet the precipitate. The samples were dried in aSavant SpeedVac Concentrator Model SVC 200H and reconstituted to theoriginal volume with cold RIA or ELISA buffer. The reconstituted sampleswere stored at -70° C. until assayed. The assay for LTB₄ was performedaccording to the directions of the [³ H]RIA kit or ELISA kit (Seragen).The total metabolite level in 1 ml of blood is calculated and reportedas ng/10⁶ neutrophils. Significance is determined by a one-way analysisof variance with LSD comparisons to control (p≦0.05). Drug effects areexpressed as percent change from control values and data for inventioncompounds is presented in Table II.

                  TABLE II                                                        ______________________________________                                        Compound of                                                                            Dose      Pretreatment                                               Example No.                                                                            mg/kg, p.o.                                                                             period (hr)                                                                              % Inhibition of LTB.sub.4                       ______________________________________                                        1        25        3          86                                              2         5        6          45                                              3        10        6          85                                              5        10        6          80                                              6        10        6          67                                              9        10        6          50                                              13        5        6          52                                              ______________________________________                                    

3. Reverse Passive Arthus Reaction

A reverse passive Arthus reaction is induced in the pleural cavity ofmale Lewis rats (150-200 g; fasted overnight prior to use) by theintravenous administration of bovine serum albumin (BSA; 4 mg/0.2 ml)followed 30 minutes later by the injection of rabbit anti-BSA (1 mg/0.2ml; lyophilized IgG fraction; Organon Teknika, West Chester, Pa.) intothe right pleural space under halothane anesthesia. Drugs or vehicle(0.5% Tween-80) control are administered orally in a volume of 1 ml/100g body weight at 1 hour prior to the anti-BSA. Animals are sacrificed ateither the time of peak eicosanoid production (i.e. 5 minutes afteranti-BSA for immunoreactive TxB₂, 10 minutes for immunoreactive LTB₄, 20minutes for immunoreactive LTC₄) or at the time of peak neutrophilinfiltration (4 hours after anti-BSA) by CO₂ inhalation. The pleuralcavity is then exposed., the fluid exudate removed by gentle vacuumaspiration and the volume of exudate is recorded. For the determinationof cellular infiltration, the pleural cavity is rinsed with 3 ml of 0.1%EDTA in sterile saline, and the recovered wash is pooled with theexudate. Cell number is determined on a model ZBI Coulter counter. Fordetermination of eicosanoid production, undiluted pleural exudate ismicrofuged and the supernatant is extracted with ethanol (8-10 timesvolume). Extracts are either stored at -20° C., or are evaporated todryness under a stream of N₂ and reconstituted in radioimmunoassay (RIA)buffer.

Eicosanoids are quantitated by RIA according to the procedure specifiedby the RIA kit manufacturer (Advanced Magnetics, Cambridge, Mass.).Briefly, 100 μl of ³ H-labeled eicosanoid and 100 μl of specificantibody are sequentially added to 100 μl of extracted pleural exudatein BGG-phosphate buffer which contains 0.01M phosphate, 0.1% bovinegamma globulin and 0.1% sodium azide at pH 7.0. Antibody-boundeicosanoid is separated from unbound eicosanoid by the addition of 750μl of dextran (0.4%)-coated charcoal (Norit A) containing 0.1% sodiumazide. The mixture is centrifuged at 2000 RPM at 5° C. for 15 minutes topellet the charcoal and adsorbed unbound eicosanoid. Antibody-boundlabeled eicosanoid is quantitated by counting in a liquid scintillationcounter, and is correlated to concentration by a standard curve.

The activity of standard drugs in this assay is as follows:

    ______________________________________                                                                           %                                                                     Dose    Inhibition                                 Antiinflammatory           mg/kg   of LTB.sub.4                               Drug       Class           p.o.    (ED.sub.50)                                ______________________________________                                        Indomethacin                                                                             NSAID; CO inhibitor                                                                           4       12                                         Naproxen                   4        0                                         Diclofenac                 10       0                                         Ketoprofen                 10      35                                         Wy-50,295-A                                                                              LO-Inhibitor    9       (15)                                       BW540C     Mixed CO/LO inhibitor   (30)                                       BW755C                             (23)                                       Phenidone                          (10)                                       ______________________________________                                    

The compounds of the invention when tested in the reverse passive Arthuspleurisy assay gave the results shown in Table III.

                  TABLE III                                                       ______________________________________                                        Compound of   Dose      % Inhibition                                          Example No.   mg/kg, p.o.                                                                             LTB.sub.4 Synthesis                                   ______________________________________                                        1             25        79                                                    2             10        59                                                    5             10        43                                                    8             10        39                                                    9             10        27                                                    13            10        17                                                    ______________________________________                                    

4. Inhibition of Brochoconstriction in Guinea Pigs Induced byExogenously Administered Antigen

Male guinea pigs (Charles River, Wilmington, Mass.) were sensitized 3-4weeks prior to antigen challenge by administration of 2 i.m. injectionsof ovalbumin, 1 into each hind limb (35 mg total). Sensitized animals(500-600 g) were fasted overnight prior to experimentation. Consciousanimals were then dosed p.o. with drug or vehicle alone (0.5% Tween 80in H₂ O) at the indicated times prior to antigen challenge, oranesthetized animals were dosed i.v. with drug or vehicle alone (DMSO) 5min prior to antigen challenge. Animals were anesthetized by urethane(2.8 g/kg i.p.). A carotid artery and jugular vein were cannulated toallow for the monitoring of blood pressure and the administration ofdrugs, respectively. The trachea was then cannulated and connected to aHarvard Apparatus rodent ventilator (S. Natick, Mass.). Spontaneousrespiration was abolished by the administration of succinylcholine (2.5mg/kg i.v.). The animals were then ventilated with room air at a rate of65 breaths per min. Airway inflation pressure was measured using aStatham pressure transducer (Gould Instruments, Cleveland, Ohio)connected to the tracheal cannula via a side-arm and recorded on a GrassInstruments recorder (Quincy, Mass.). The tidal volume (approximately 10cc/kg) was adjusted to give a baseline inflation pressure of 8-10 cm H₂O at end inspiration. Animals were then allowed 20 min to stabilize.

Following the stabilization period, animals were given i.v. injectionsof pyrilamine (5 mg/kg), propranolol (0.1 mg/kg) and indomethacin (10mg/kg) at 15, 10 and 5 min, respectively, prior to antigen challenge.This pretreatment results in an LT-dependent bronchoconstrictionfollowing antigen challenge, which was accomplished by i.v.administration of ovalbumin (10 mg/kg). Only one bronchoconstriction peranimal was induced. End-inspiratory inflation pressure (in cm H₂ O overbaseline) was measured at 5 min post-antigen challenge. A mean value andstandard error for the % inhibition of control bronchoconstriction ineach drug-treated group was then calculated. Data for inventioncompounds are presented in Table IV.

                  TABLE IV                                                        ______________________________________                                        Compound of                                                                            Dose      Administration                                                                            Bronchoconstriction                            Example No.                                                                            (mg/kg)   Route       % Inhibition                                   ______________________________________                                        3        10        i.v.        84                                             3        25        p.o.        26                                             4        10        i.v.        53                                             5        25        p.o.        29                                             6        10        i.v.        70                                             7        10        i.v.        58                                             7        25        p.o.        18                                             8        10        i.v.        41                                             9        10        i.v.        65                                             13       10        i.v.        29                                             ______________________________________                                    

5. Inhibition of Copper Ion Mediated Oxidation of Low DensityLipoprotein

In this in vitro assay, the inhibition of Cu⁺² mediated oxidation ofrabbit or monkey LDL by an invention compound is determinedspectrophotometricallly. Oxidation of LDL results in the formation ofLDL-diene which absorbs light at 532 nm. Inhibition of oxidation of LDLleads to a decrease in absorbance at 532 nm.

Rabbit or monkey LDL is prepared according to the procedures of Havel,Eder and Gragdon, "The Distribution and Chemical Composition ofUltracentrifugally Separated Lipoproteins in Human Serum,: J. Clin.Invest. 34, 1345-1353 (1955) and Parhtasarathy, Wieland and Steriberg,"A Role for Endothelial Cell Lipoxygenase in the Oxidative Modificationof Low Density Lipoprotein," Proc. Natl. Acad. Sci. USA 86, 1046-1050(1989). Test compound solutions are prepared by dissolving the inventioncompounds in ethanol at concentrations up to 248 μM. The medium used isDulbecco's phosphate buffered saline containing 0.5 mg/ml bovine serumalbumin. For standards, 0 to 10 μl of an aqueous solution of1,1,3,3-tetraethoxypropane (1 μmol/ml H₂ O) in 4.1 ml of medium is used.

Test compound solution (100 μl) is added to 4 ml of medium in incubationtubes. To each tube is added 10 μl of LDL solution and 25 μl of aqueouscopper sulfate solution (1.32 mg/ml H₂ O). The tubes are incubated at37° C. for 90 minutes and the oxidation reaction quenched by addition of1 ml of thiobarbituric acid solution (0.67% in 50% acetic acid). Thetubes are heated at 90° C. for 1 hour, then chilled in an ice bath andthe chromophore extracted into 2 ml of n-butanol. Absorbence is read at532 nm and the results are reported as nmols of malondialdehydeequivalents.

Significant differences (p<0.05) are determined by the Dunnett T-test orby the Student-Newman-Keuls Test for significant differences betweenmeans. The assay is conducted using several concentrations [I] of theinhibitor test compounds. The LDL solution concentrations at differentexperiments were either 1.5 mg/ml, 2.5 mg/ml or 10.3 mg/ml. The IC₅₀ isdetermined by non-linear regression, plotting log [I] vs. % inhibition.(Reference: K. Yagi, Biochemical Medicine 15,212-216 (1976)). Theresults obtained with invention compounds are shown in Table V.

                  TABLE V                                                         ______________________________________                                        Example No.    IC.sub.50 (μM)                                              ______________________________________                                        1              0.64                                                           6              0.59                                                           9              0.57                                                           ______________________________________                                    

Pharmaceutical Composition

When the compounds of the invention are employed in the treatment ofallergic airway disorders, inflammation, or atherosclerosis, they can beformulated into oral dosage forms such as tablets, capsules and thelike. The compound can be administered alone or by combining them withconventional carriers, such as magnesium carbonate, magnesium stearate,talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth,methylcellulose, sodium carboxymethylcellulose, low melting wax, cocoabutter and the like. Diluents, flavoring agents, solubilizers,lubricants, suspending agents, binders, tablet-disintegrating agents andthe like may be employed. The compounds may be encapsulated with orwithout other carders. In all cases, the proportion of activeingredients in said compositions both solid and liquid will be at leastto impart the desired activity thereto on oral administration. Thecompounds may also be administered parenterally, in which case they areused in the form of a sterile solution containing other solutes, forexample, enough saline or glucose to make the solution isotonic. Foradministration by inhalation or insufflation, the compounds may beformulated into an aqueous or partially aqueous solution, which can thenbe utilized in the form of an aerosol.

The dosage requirements vary with the particular compositions employed,the route of administration, the severity of the symptoms presented andthe particular subject being treated. Treatment will generally beinitiated with small dosages less than the optimum dose of the compound.Thereafter the dosage is increased until the optimum effect under thecircumstances is reached. In general, the compounds of the invention aremost desirably administered at a concentration that will generallyafford effective results without causing any harmful or deleterious sideeffects, and can be administered either as a single unit dose, or ifdesired. the dosage may be divided into convenient subunits administeredat suitable times throughout the day.

What is claimed is:
 1. A method of treating atherosclerosic plaqueformation in a mammal in need thereof which comprises administration ofa therapeutically effective amount of a compound having the formula:##STR13## wherein R¹ and R³ are independently hydrogen, fluorine,chlorine, bromine, iodine, C₁ -C₆ alkyl, trifluoromethyl, C₁ -C₆ alkoxy,or C₁ -C₆ trifluoroalkoxy; R² and R⁴ are hydrogen or methylindependently;R⁵ is hydrogen, methyl or hydroxy; X and Z areindependently oxygen or sulfur; and Y is --CH₂ --, --CH(CH₃)--, or--CH═CHCH(CH₃)--.